Pollution by diazepam (DZP) is increasingly recognized as a major threat to aquatic organisms, but knowledge about its potential risk to fish is still limited. In this study, we exposed female and male Japanese medaka (Oryzias latipes) to environmentally relevant DZP (0.8 and 8 µg/L) for 28 days and investigated variation in their behavior (on days 7, 14, and 28) and brain neurotransmitter levels (on day 28). The results showed that DZP could be accumulated in the brain and gonads in Japanese medaka. When two fish of the same sex were placed in an aquarium, DZP exposure exhibited typical sedative effects on females (on day 7) and males (on days 7 and 14). However, these sedative effects on both sexes were no longer present after 28 days of exposure. Exposure to DZP induced sex-specific impacts on the social interactions of medaka on days 7, 14, and 28 of exposure in a time-dependent manner. When both sexes were placed into an aquarium in a ratio of 1:1, DZP could significantly alter their locomotor activity and social interaction on days 14 and 28 of the exposure. After 28 days of exposure, DZP significantly altered the levels of several neurotransmitters in the brain of medaka, also in sex-specific manners. The alterations in dopamine and serotonin levels exhibited significant correlations with the increased social interaction between females. At the same time, that of γ-aminobutyric acid significantly correlated to the decreased social interaction between males. Our findings suggest that chronic exposure to DZP, even at environmentally relevant concentrations, can accumulate in the brains and gonads of fish, and alter their behaviors by mediating brain neurotransmitter levels, which may further disturb their reproduction and population dynamics.
Keywords: Diazepam; Japanese medaka; Neurotransmitters; Sex-specific impacts; Social interaction.
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